Corneal wound healing without scarring regenerates corneal transparency. Corneas that heal with scarring have opacities and obstructed vision. Thus, understanding the pathways that influence regenerative rather than fibrotic healing is an essential step towards therapeutically promoting healthy repair. Upon corneal wounding, the normally quiescent cells in the stroma differentiate into fibroblasts, which secrete proteases as they migrate. This proposal is focused on the contributions of the extracellular serine protease, urokinase-type plasminogen activator (uPA), which is expressed by corneal fibroblasts after wounding. When uPA binds to its cell-surface receptor, uPAR, uPA generates plasmin at the cell-matrix interface. Plasmin degrades extracellular matrix (ECM) and activates latent growth factors such as TGF?. TGF? stimulates fibroblast proliferation and migration, and induces the differentiation of motile fibroblasts into non-motile myofibroblasts, which are essential for matrix contraction and wound closure. One way in which TGF? may be eliciting its dual effects is through the induction of plasminogen activation inhibitor (PAI-1) expression. There is a significant body of data that the opposing effects of PAI-1 are concentration dependent: low concentrations of PAI-1 induce cell proliferation and migration, whereas, high concentrations inhibit these processes. The hypothesis being tested in this proposal is that after corneal wounding, local PAI-1 concentrations are mediated by binding to Vn. Further, that low concentrations of PAI-1 result in corneal fibroblast proliferation and migration whereas high concentrations of PAI-1 induce uPA/uPAR downregulation and result in myofibroblast differentiation.
The specific aims are to determine if 1) TGF? concentration regulates PAI-1 levels, uPA activity, and myofibroblast differentiation. 2) Vn mediates the effects of TGF? and PAI-1 on uPA activity, cell migration, and myofibroblast differentiation. 3) Vn, PAI-1 and integrin expression change throughout wound healing with changing stromal phenotypes. 4) uPAR cleavage into a non-uPA binding form inhibits cell migration and induces myofibroblast differentiation. The results of these studies will lead to a better understanding of the mechanisms that guide regenerative wound repair.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY017030-04
Application #
7626280
Study Section
Anterior Eye Disease Study Section (AED)
Program Officer
Shen, Grace L
Project Start
2006-06-01
Project End
2011-05-31
Budget Start
2009-06-01
Budget End
2010-05-31
Support Year
4
Fiscal Year
2009
Total Cost
$329,169
Indirect Cost
Name
Icahn School of Medicine at Mount Sinai
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
078861598
City
New York
State
NY
Country
United States
Zip Code
10029
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Wang, Lingyan; Pedroja, Benjamin S; Meyers, Erin E et al. (2012) Degradation of internalized ?v?5 integrin is controlled by uPAR bound uPA: effect on ?1 integrin activity and ?-SMA stress fiber assembly. PLoS One 7:e33915
Wang, Lingyan; Ko, Chun-Ying; Meyers, Erin E et al. (2011) Concentration-dependent effects of transforming growth factor ?1 on corneal wound healing. Mol Vis 17:2835-46
Stepp, Mary Ann; Daley, William P; Bernstein, Audrey M et al. (2010) Syndecan-1 regulates cell migration and fibronectin fibril assembly. Exp Cell Res 316:2322-39
Tall, Edward G; Bernstein, Audrey M; Oliver, Noelynn et al. (2010) TGF-?-stimulated CTGF production enhanced by collagen and associated with biogenesis of a novel 31-kDa CTGF form in human corneal fibroblasts. Invest Ophthalmol Vis Sci 51:5002-11
Pedroja, Benjamin S; Kang, Leah E; Imas, Alex O et al. (2009) Plasminogen activator inhibitor-1 regulates integrin alphavbeta3 expression and autocrine transforming growth factor beta signaling. J Biol Chem 284:20708-17